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1.
Figure 2

Figure 2. From: Distribution, persistence and efficacy of adoptively transferred central and effector memory-derived autologous SIV-specific CD8+ T-cell clones in rhesus macaques during acute infection.

TCM- and TEM-derived CD8+ T-cell clones exhibit similar functional reactivity and both display an effector memory surface phenotype after culture in vitro. TCM- and TEM-derived SIV-specific CD8+ T-cell clones from two rhesus macaques, AZ15 and DBN2, were stimulated with PBMC pulsed with the appropriate peptides (A) or autologous SIV-infected CD4+ T-cell clones (B). IFN-γ-expression by the TCM- and TEM-derived clones was measured by ICS and flow cytometry. The surface phenotype of TCM- and TEM-derived clones was determined using flourochrome conjugated mAbs to CD28 and CD95 followed by flow cytometric analyzes (C).

Jacob T. Minang, et al. J Immunol. ;184(1):315-326.
2.
Figure 1

Figure 1. From: Distribution, persistence and efficacy of adoptively transferred central and effector memory-derived autologous SIV-specific CD8+ T-cell clones in rhesus macaques during acute infection.

Gating strategy and flow cytometry sorting analysis of central (TCM) and effector (TEM) memory CD8+ T-cell populations from a rhesus macaque (DBN2) immunized with a DNA vaccine construct containing full length SIVmac239 Gag, Pol, Vif, Tat, and Nef sequences. CD8+ lymphocytes were singlet-gated and defined as memory cells based on surface expression of CD95. TCM and TEM fractions were further defined and sorted based on CD28 and CCR7 expression.

Jacob T. Minang, et al. J Immunol. ;184(1):315-326.
3.
Figure 7

Figure 7. From: Distribution, persistence and efficacy of adoptively transferred central and effector memory-derived autologous SIV-specific CD8+ T-cell clones in rhesus macaques during acute infection.

Escape mutation in the SIV Nef159-167YY9 epitope detected <3 weeks post infection. Virus isolated from plasma collected from the Mamu A*02-positive monkey AZ15 on days 12 and 20 after challenge with SIVmac239 (days 9 and 17 post T cell infusion) was analyzed by direct sequencing spanning the SIV Nef YY9 epitope. The nucleotide change is highlighted by black circles with the encoded amino acid indicated.

Jacob T. Minang, et al. J Immunol. ;184(1):315-326.
4.
Figure 3

Figure 3. From: Distribution, persistence and efficacy of adoptively transferred central and effector memory-derived autologous SIV-specific CD8+ T-cell clones in rhesus macaques during acute infection.

Intravenously infused ex vivo expanded virus-specific TCM- and TEM-derived CD8+ T-cell clones show similar distribution and persistence in vivo. Half of the clonal population of TCM- and TEM-derived virus-specific CD8+ T-cell clones from two rhesus macaques, DBN2 and AZ15, were stained with PKH26 and CFSE, respectively, and the pool of stained and unstained cells adoptively transferred to the monkeys. The distribution and persistence of the infused cells was analyzed by flow cytometry by gating for the CD3+CD8+ T-cell fraction of the “live gate” of the forward and side scatter plot (A). PBMC (B), LN and GALT (C) samples were analyzed at indicated time points post infusion.

Jacob T. Minang, et al. J Immunol. ;184(1):315-326.
5.
Figure 6

Figure 6. From: Distribution, persistence and efficacy of adoptively transferred central and effector memory-derived autologous SIV-specific CD8+ T-cell clones in rhesus macaques during acute infection.

Adoptive transfer of virus-specific CD8+ T cells does not significantly affect the endogenous SIV-specific T-cell response. (A) PBMC collected at the indicated time points from the two Mamu A*01 positive monkeys, DBN2 and DBK1 were stained with SIV Tat SL8 or Gag CM9 tetramers and CD8 mAb. (B) PBMC from DBN2 and DBK1 collected pre- and post- infusion were stimulated with SIV Tat SL8, Gag CM9 and Acc peptide pool and the percentage of CD8+ IFN-γ-producing T cells determined by flow cytometry, or (C) the number of IFN-γ-producing cells determined by ELISpot. (D) PBMC from AZ15, DBN2 and DBK1 collected on day 39 post infusion were stimulated with SIV Acc, Env, Gag and Pol peptide pools and the number of antigen-induced IFN-γ-producing T cells determined by ELISpot.

Jacob T. Minang, et al. J Immunol. ;184(1):315-326.
6.
Figure 5

Figure 5. From: Distribution, persistence and efficacy of adoptively transferred central and effector memory-derived autologous SIV-specific CD8+ T-cell clones in rhesus macaques during acute infection.

T cell persistence, function and viral load in BAL samples. (A) Virus-specific TCM- and TEM-derived CD8+ T-cell clones from two rhesus macaques, DBN2 and AZ15, half of which were stained with PKH26 and CFSE, respectively, were infused 3 days after challenge with SIVmac239. Infused cells were tracked using flow cytometry at indicated time points (B). Cell-free viral RNA and cell-associated viral DNA and RNA were determined at indicated time-points by QPCR. DBN2 and AZ15 received virus-specific CD8+ T cells. Grey dashed lines show threshold of detection of cell-associated viral DNA and RNA (>10 copies/100,000 cells). (C) Cells from BAL from DBN2 were stimulated with SIV Acc peptide pool or the SIV Gag CM9 peptide as indicated. The frequency of IFN-γ-expressing cells was determined by ICS and flow cytometry.

Jacob T. Minang, et al. J Immunol. ;184(1):315-326.
7.
Figure 4

Figure 4. From: Distribution, persistence and efficacy of adoptively transferred central and effector memory-derived autologous SIV-specific CD8+ T-cell clones in rhesus macaques during acute infection.

Adoptive transfer of ex vivo expanded virus-specific CD8+ T-cell clones show no measurable effect on the virus load or the number of circulating CD4+ T cells in peripheral blood. Virus load in two animals infused with virus-specific CD8+ T cells, DBN2 and AZ15, as well as a control animal, DBK1, was determined on day 0, 3 (day of infusion), 4, 5 and 7 post challenge and once a week thereafter, by analyzing cell-free plasma viral RNA (A) or cell-associated viral DNA (B) levels by QPCR. The frequency of circulating CD4+ T cells in PBMC was determined using the BD Tru Count™ kit (C).

Jacob T. Minang, et al. J Immunol. ;184(1):315-326.

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